CST, 32578T, DNMT3A (D2H4B) Rabbit Monoclonal Antibody

Monoclonal Antibody for studying DNMT3A. Validated for WB,IP,IF,F,ChIP,ChIP,C&R,C&T. Available in 2 sizes. Highly specific and rigorously validated in-house, DNMT3A (D2H4B) Rabbit Monoclonal Antibody (CST #32578) is ready to ship. Product Usage Information For optimal ChIP and ChIP-seq results, use 10 μl of antibody and 10 μg of chromatin (approximately 4 x 10 6 cells) per IP. This antibody has been validated using SimpleChIP ® Enzymatic Chromatin IP Kits. The CUT&RUN dilution was determined using CUT&RUN Assay Kit # 86652 . The CUT&Tag dilution was determined using CUT&Tag Assay Kit # 77552 . Western Blotting: 1:1000 Immunoprecipitation: 1:50 Immunofluorescence (Immunocytochemistry): 1:800 - 1:1600 Flow Cytometry (Fixed/Permeabilized): 1:1600 Chromatin IP: 1:50 Chromatin IP-seq: 1:50 CUT&RUN: 1:50 CUT&Tag: 1:50 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunofluorescence (Immunocytochemistry), Flow Cytometry (Fixed/Permeabilized), Chromatin IP, Chromatin IP-seq, CUT&RUN, CUT&Tag Specificity / Sensitivity DNMT3A (D2H4B) Rabbit Monoclonal Antibody recognizes endogenous levels of total DNMT3A protein. This antibody detects multiple isoforms of DNMT3A, including isoform 1 and isoform 2. This antibody does not cross-react with DNMT3B or other DNMT proteins. Species Reactivity: Human Source / Purification Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro233 of human DNMT3A protein. Background Methylation of DNA at cytosine residues in mammalian cells is a heritable, epigenetic modification that is critical for proper regulation of gene expression, genomic imprinting and development (1,2). Three families of mammalian DNA methyltransferases have been identified: DNMT1, DNMT2, and DNMT3 (1,2). DNMT1 is constitutively expressed in proliferating cells and functions as a maintenance methyltransferase, transferring proper methylation patterns to newly synthesized DNA during replication. DNMT3A and DNMT3B are strongly expressed in embryonic stem cells with reduced expression in adult somatic tissues. DNMT3A and DNMT3B function as methyltransferases that methylate previously unmethylated regions of DNA. DNMT2 is expressed at low levels in adult somatic tissues and its inactivation affects neither nor maintenance DNA methylation. DNMT1, DNMT3A, and DNMT3B together form a protein complex that interacts with histone deacetylases (HDAC1, HDAC2, Sin3A), transcriptional repressor proteins (RB, TAZ-1), and heterochromatin proteins (HP1, SUV39H1) to maintain proper levels of DNA methylation and facilitate gene silencing (3-8). Improper DNA methylation contributes to diseased states such as cancer (1,2). Hypermethylation of promoter CpG islands within tumor suppressor genes correlates with gene silencing and the development of cancer. In addition, hypomethylation of bulk genomic DNA correlates with and may contribute to the onset of cancer. DNMT1, DNMT3A, and DNMT3B are overexpressed in many cancers, including acute and chronic myelogenous leukemias, in addition to colon, breast, and stomach carcinomas (9-12). Alternate Names DNA (cytosine-5-)-methyltransferase 3 alpha; DNA (cytosine-5)-methyltransferase 3A; DNA cytosine methyltransferase 3A2; DNA methyltransferase 3 alpha; DNA methyltransferase HsaIIIA; DNA MTase HsaIIIA; DNM3A; DNMT3A; DNMT3A2; HESJAS; M.HsaIIIA; TBRS Specification REACTIVITY: H SENSITIVITY: Endogenous MW (kDa): 130, 95, 85 Source/Isotype: Rabbit IgG